Recombination-mediated PCR-directed plasmid construction in vivo in yeast

Nucleic Acids Res. 1997 Jan 15;25(2):451-2. doi: 10.1093/nar/25.2.451.


We have extended the technique of PCR-directed recombination in Saccharomyces cerevisiae to develop a simple method for plasmid or gene construction in the absence of suitable restriction sites. The DNA to be cloned is PCR-amplified with 30-40 bp of homology to a linearized yeast plasmid. Co-transformation into yeast results in homologous recombination at a position directed by the PCR oligonucleotides.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cloning, Molecular / methods
  • DNA Primers
  • DNA, Recombinant / genetics
  • Genes, Fungal
  • Genes, Reporter / genetics
  • Green Fluorescent Proteins
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Plasmids / genetics
  • Plasmids / metabolism*
  • Polymerase Chain Reaction
  • Recombination, Genetic / genetics*
  • Saccharomyces cerevisiae / genetics*
  • Transformation, Genetic / genetics


  • DNA Primers
  • DNA, Recombinant
  • Luminescent Proteins
  • Green Fluorescent Proteins